• Title/Summary/Keyword: 변성 구배 젤 전기영동

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Prokaryotic Communities of Halophilic Methylotrophs Enriched from a Solar Saltern (염전으로부터 농화배양된 호염 메틸영양미생물 군집의 특성)

  • Kim, Jong-Geol;Park, Soo-Je;Rhee, Sung-Keun
    • Korean Journal of Microbiology
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    • v.46 no.3
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    • pp.286-290
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    • 2010
  • C-1 compounds are observed in anaerobic sediment of high salt environments. Thus, surface sediments and waters from these environments are therefore potential habitats for aerobic methylotrophic microorganisms. The soil samples collected from saltern and tidal flat as inoculums and methanol as carbon and energy source was supplied. After subculture depending on the salt concentration, methanol oxidizing bacteria growth condition investigated, the results of methanol oxidizing bacteria can grow in salt conditions, and the maximum concentration was 20%. Analysis based on denaturing gradient gel electrophoresis of 16S rRNA genes indicates that Methelyophaga-like bacteria were dominants of methylotrophs in the enrichment culture. Quantitative PCR showed that archaeal cells were about 1-10% of bacterial cells. Additionally archaea were assumed not to be involved in methanol oxidation since bacterial antibiotics completely blocked the methanol oxidation. Our results suggest that Methelyophaga-like bacteria could be involved in C-1 compounds oxidation in hypersaline environments although those activities are sensitive to salinity above 20%.

Korean Paddy Soil Microbial Community Analysis Method Using Denaturing Gradient Gel Electrophoresis (Denaturing gradient gel electrophoresis를 이용한 한국의 논 토양 미생물 다양성 분석 방법)

  • Choe, Myeongeun;Hong, Sung-Jun;Lim, Jong-Hui;Kwak, Yunyoung;Back, Chang-Gi;Jung, Hee-Young;Lee, In-Jung;Shin, Jae-Ho
    • Journal of Applied Biological Chemistry
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    • v.56 no.2
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    • pp.95-100
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    • 2013
  • Soil microbes are important integral components of soil ecosystem which have significant and diverse role in organic matter decomposition, nitrogen cycling, and nitrogen fixation. In this study an effective denaturing gradient gel electrophoresis (DGGE) method was employed for paddy soil microbial diversity survey. For optimum paddy soil microbial DNA extraction, different methods such as Lysis buffer, skim milk bead, sodium phosphate buffer, Epicentre Soil Master DNA extraction kit (Epicentre, USA) and Mo Bio Power Soil DNA kit (MO BIO, USA) methods were utilized. Among all the method, using Mo Bio Power Soil kit was most effective. DGGE analysis of Bacteria was carried out at 6% polyacylamide gel and 45-60% denaturing gradient in the optimal conditions. Whereas DGGE analysis of fungi was done at 6% polyacrylamide gel and 45-80% denaturing gradient in the optimal conditions. By applying the above assay, it was found that variation within the microbial community of paddy soil occurs by a factor of time. DGGE assay used in this study through for a variety of soil microbial analysis suggests the potential use of this method.